Recent investigations on plant molybdenum-containing enzymes that include xanthine dehydrogenase (EC 1.1.1.204) and xanthine oxidase (EC 1.1.3.22), nitrate reductase (EC 1.7.1.1-3), aldehyde oxidase (EC 1.2.3.1), and sulfite oxidase (EC 1.8.3.1) are reviewed. The enzymes belong to closely related protein family and share common structural features. Special attention is being paid to the recently solved crystal structures their implications for the substrate binding and catalytic mechanism.

A field study was conducted to evaluate the drought tolerance of three sorghum [Sorghum bicolor (L.) Moench] cultivars, Gadambalia, Arous elRimal and Tabat, and quantify the physiological bases for differences in their drought tolerance. Water stress reduced shoot dry mass of Gadambalia, Arous elRimal and Tabat by 43, 46 and 58 %, respectively. The respective reduction in leaf area of the three cultivars was 28, 54 and 63 %. The reduction in net photosynthetic rate, stomatal conductance and transpiration rate due to water stress was lowest in Gadambalia and highest in Tabat. The leaf water potentials and relative water contents of Gadambalia under wet and dry treatments were similar, while those of Tabat were significantly reduced by water stress. The lowest and highest liquid water flow conductance was displayed by Tabat and Gadambalia, respectively. Drought tolerance in Gadambalia is associated with its smaller leaf area, higher liquid water flow conductance, and ability to maintain high leaf water potential, relative water content, stomatal conductance, transpiration rate and photosynthetic rate under drought stress.

In vitro regeneration of Melia azedarach L. was studied. Shoots were regenerated from calli initiated from leaflets of in vitro growing plants. The best medium for establishment of cultures was Murashige and Skoog (MS) medium with 4.44 μM benzylaminopurine (BAP) + 0.46 μM kinetin (KIN) + 16.29 μM adenine sulphate (ADE). Regenerated shoots were multiplied in MS + 0.44 μM BAP + 0.37 μM KIN + 3.26 μM ADE. Maximal rooting of 89 % was achieved by culture of regenerated shoots in MS + 12.26 μM indole-3-butyric acid for 3 d and subsequently in MS lacking growth regulators for 27 d. Rooted shoots were acclimatized and successfully transferred to soil.

A micro-propagating system based on the young stem node segments of Sophora flavescens Ait. (Fabaceae) was established. Murashige and Skoog (MS) basal medium supplemented with 8.88 μM 6-benzyladenine (BA) plus 2.69 μM α-naphthalene acetic acid (NAA) and that with only 5.37 μM NAA were found the best in promoting proliferation of shoots and induction of root, respectively. Percentages of shoot induction and number of shoot per explant were up to 93.4 % and 4.2 and rooting rate to 82.4 %, respectively. The segments of the regenerated shoots could be continuously induced to reproduce new shoots through subculture on the same medium in 30-d intervals and still kept this activity after being subcultured for 6 generations. After the regenerated plantlets were transplanted to field, they grew well, showing no any visible abnormalities.

Dimorphic seeds of Atriplex prostrata were removed from cold dry storage monthly over a one year period to test for fluctuations in seed dormancy and germination rate. For each seed type, four replicates of 25 seeds were exposed to four alternating night/day temperature regimes mimicking seasonal fluctuations in Ohio: 5/15 °C; 5/25 °C; 15/25 °C and 20/35 °C with a corresponding 12-h photoperiod (20 μmol m^-2 s^-1; 400 - 700 nm). We found a significant three-way interaction of seed size, temperature and month for both percent germination and the rate of germination. Large seeds showed the greatest germination at the 20/35 °C and 5/25 °C temperature regimes and small seeds at the 5/25 °C regime. Large seeds had greater germination at all temperatures as compared to small seeds. Large seeds had the fastest germination rates at 20/35 °C followed by 5/25 °C whereas small seeds had the fastest rates at 5/25 °C followed by 20/35 °C.

Overexpression of antifungal pathogenesis-related (PR) proteins in crop plants has the potential for enhancing resistance against fungal pathogens. Thaumatin-like proteins (TLPs) are one group (PR-5, permatins) of antifungal PR-proteins isolated from various plants. In the present study, a plasmid containing a cDNA of rice tlp (D34) under the control of the CaMV-35S promoter was introduced into tobacco plants through Agrobacterium-mediated transformation system. A considerable overproduction of TLP was observed in transformed tobacco plants by Western blot analysis. There was a large accumulation of tlp mRNA in transgenic plants as revealed by Northern blot analysis. Southern blot analysis of the DNA from transgenic tobacco plants confirmed the presence of the rice tlp gene in the genomic DNA of transgenic tobacco plants. Immunoblot analysis of intracellular and extracellular proteins of transgenic tobacco leaves using a Pinto bean TLP antibody demonstrated that the 23-kDa TLP was secreted into the extracellular matrix. T₂ progeny of regenerated plants transformed with TLP gene were tested for their disease reaction to Alternaria alternata, the brown spot pathogen. Transgenic tobacco plants expressing TLP at high levels showed enhanced tolerance to necrotization caused by the pathogen.

In the present study three species of Heritiera Aiton (Sterculiaceae) were characterized using 9 amplified fragment length polymorphism (AFLP) primer combinations and the genetic relationship among these three species was assessed, Nine AFLP primer combinations yielded 445 bands out of which 210 were monomorphic and 235 were polymorphic. Out of the 235 polymorphic bands 79 were present only in a single species. Among the total amplified bands 255 were shared between H. fomes and H. littoralis, 225 were shared between H. fomes and H. macrophylla and 306 bands were shared between H. littoralis and H. macrophylla. The cluster analysis showed H. littoralis is closer to H. macrophylla than H. fomes. The similarity between H. fomes and H. littoralis was higher than that of H. fomes and H. macrophylla. The present study indicates that H. littoralis is better classified as mangrove associate or back mangal than a true mangrove.

Effective protocol was established for micropropagation of the medicinal plant Eupatorium triplinerve Vahl through rapid axillary bud proliferation and ex vitro rooting. Murashige and Skoog (MS) medium fortified with 8.87 μM benzylaminopurine (BAP) and 2.46 μM indole-3-butyric acid (IBA) was the best for axillary bud proliferation and developed a mean of 8.1 shoots per node. Excision and culture of the node segments of the in vitro shoots on medium supplemented with the same concentration of growth regulators developed more than 30 shoots within 40 d. Shoot multiplication did not exhibit decrease in the number of shoots even at 7^th subculture. Dipping of the basal end of shoots in 2.46 μM IBA solution for 10 d induced roots and its transfer to small pots facilitated the survival of all rooted shoots (100 %). Ex vitro rooting by direct transfer of the shoots from multiplication medium showed 92 % survival.

The experiments were carried out with germinating maize seeds (Zea mays L.), grown 6 d in the dark at 26°C. Before germination the seeds were soaked for 4 h in solutions containing 1 mM abscisic acid (ABA), 0.1 mM N₁-(2-chloro-4-pyridyl)-N₂ phenylurea (4PU-30) and their combination. The influence of plant growth regulators on the length, fresh (FM) and dry (DM) masses, proteolytic activities and soluble protein fractions in shoots, roots and endosperm were studied. As compared to control the seedlings treated with ABA showed lower length, FM and DM of shoots and roots, and lower proteolytic activities. As a consequence of suppression of both growth and protein breakdown, these seedlings possessed higher protein content in endosperm. 4PU-30 partially decreased the ABA suppressing effects.

Exogenously applied methyl jasmonate (MeJA) might induce the formation of necrotic lesions that closely resemble hypersensitive response lesions. Cellular damage, restricted to the infiltrated zone, was accompanied with the production of H₂O₂ from the oxidative burst. H₂O₂ generated in response to MeJA can be histochemically detected in cells surrounding the necrotic lesions as well as in the vascular tissues. The response is systemic and maximizes with time. Among 12 plant species from different families that were assayed for both hypersensitive reaction (HR)-like response and H₂O₂ generation, only woody species exhibited both MeJA-inducible HR cell death and the generation of H₂O₂. To assess the role of H₂O₂ in MeJA-induced HR-like cell death, a gain and loss of function strategy was employed. The cumulative results indicate that H₂O₂ is neither necessary nor sufficient for MeJA-inducible cell death and that O₂^- rather than H₂O₂ might be responsible.

Zinc adsorption by two green algae, Scenedesmus obliquus and Scenedesmus quadricauda, was investigated. The maximum adsorption capability of zinc ion obtained from the Langmuir adsorption isotherms was higher for S. obliquus (6.67) than for S. quadricauda (5.03), and S. obliquus was more tolerant to zinc phytotoxicity than S. quadricauda. Lower concentrations of zinc increased dry mass, chlorophylls a and b, carotenoids, and total amino acid contents in both algae. On the other hand, higher concentrations of zinc were inhibitory for growth and the other metabolic activities in a concentration dependent manner.

A protocol was developed for direct differentiation of multiple shoot buds from leaf explants of Cajanus cajan. In a modified Murashige and Skoog's medium supplemented with 2.22 µM benzyladenine (BA), 0.57 µM indole-3-acetic acid (IAA) and 41 µM adenine sulphate (AdS), the segments of basal halves of the first two leaves of a young seedling incubated on filter paper bridges in liquid medium took 20 - 25 d to differentiate shoot buds. The explants after transfer to solidified medium, with lower concentration of BA (0.22 μM) resulted in fast growing healthy shoots. The developed shoots (measuring ca. 3 cm) were rooted in a medium supplemented with 1.42 µM IAA. They were subsequently grown in pots with soil with more than 80 % transplantation success.

The effects of increasing relative humidity on the growth and salt tolerance of two melon (Cucumis melo L.) cultivars, Revigal C-8 (salt sensitive) and Galia (salt tolerant) was investigated. One month after germination, the plants were exposed for 15 d to 0 (control) and 80 mM NaCl, under relative humidity (RH), 30 and 70 %. The growth of the whole plant, leaf, stem and root of cv. Revigal C-8 was increased with increasing RH. On the other hand, cv. Galia showed an increase in root growth with increasing RH only under the NaCl treatment. Under salinity, most of the Na⁺ was withheld in the stems. An increase in RH in the NaCl treatment significantly decreased Na⁺ and Cl^- concentrations in leaves of cv. Revigal C-8, while it had no effect on their concentrations in cv. Galia. In both cultivars, increasing RH under NaCl condition significantly decreased water contents in leaves and stems, and increased osmotic potential in roots. The amount of the root exudate of cv. Galia was significantly decreased with increasing RH, while it was not affected in cv. Revigal C-8. Under the NaCl treatment, cv. Galia had significantly higher leaf osmotic potential than cv. Revigal C-8 at both relative humidities and higher amount of root exudate at 30 % RH.

Direct somatic embryogenesis in celandine (Chelidonium majus L.) was achieved in epicotyl explants of seedlings after prolonged cultivation on Murashige and Skoog medium with or without plant growth regulators. Somatic embryos developed into plantlets which entered additional cycles of somatic embryogenesis. Cultures consisting of plantlets with prolonged embryogenic potential were maintained for five years on plant growth regulator free medium. Embryos which developed into rooted plantlets could be acclimated in a glasshouse enabling thus a continuous propagation scheme to be established.

The ratio of activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase (G6P DH/6PG DH), and the contents of glucose-6-phosphate (G6P), 6-phosphogluconate (6PG) and fructose-6-phosphate (F6P) were studied at various stages of potato virus Y (PVY) multiplication in Nicotiana tabacum cv. Samsun. G6P DH/6PG DH increased through the experiment from 0.42 to 0.53 in leaves of healthy tobacco, and up to 0.59 in PVY systemically infected leaves. However, these ratios in the ruptured protoplast preparations, and the chloroplast and cytosol fractions of healthy protoplasts were similar to that from infected ones. The ratio lower than 1, found in the healthy and/or PVY- infected leaf tissues and in the infected protoplasts as well, confirms the assumption that G6P DH is the control enzyme of oxidative pentosephosphate pathway not only in the healthy but also in the infected plants. The contents of G6P, 6PG and F6P in the period of the highest PVY multiplication were strongly decreased (to 30 - 50 % when compared with control healthy leaves) and were negatively correlated with the G6P DH and 6PG DH activities.

The aim of this research was to set up a regeneration protocol from mature explants of Prunus avium L. cv. Hedelfinger and to develop an early screening method for selection of putative somaclones based on morphological and physiological traits regulated by the spectral quality of radiation. DNA analyses of a new somaclone named HS, conducted using Inter Simple Sequence Repeat (ISSR), revealed a polymorphism between the somaclone HS and wild type propagated by microcuttings. When grown under different spectral quality of radiation, somaclone HS showed a different pattern of growth and development compared to the wild type with the main modifications related to apical dominance and chlorophyll production. Somaclone HS showed reduced apical dominance compared to the wild type. Wild type shoots grown in darkness showed chlorophyll a and b contents at levels in both cases comparable to those recorded under red radiation while HS did not retain the same capability.

The study was focused on changes of anatomical and histochemical parameters of buds of 4-year-old Norway spruce (Picea abies L. Karst) trees subjected to simulated acid rain (SAR). Solutions of pH 2.9 and 3.9 were applied by spraying on shoot and/or by watering for two years. No macroscopic changes of buds or needles were observed in connection with SAR application and the only induced change was 2-week earlier onset of bud break in all treated variants compared to the control. Two-year treatment caused decrease in number of leaf primordia and increase in number of living bud scales in treated dormant buds while these parameters remained unchanged in the control buds. Treatments with solution of pH 2.9 caused decrease of flatness of bud apical meristem during the vegetative season. Increased activity of non-specific esterase in treated buds occurred during dormancy and bud break and the enhanced accumulation of phenolic compounds was detected at the beginning of shoot growth. Changes in histochemical parameters of bud tissues were induced mainly by spraying of shoots and can thus be qualified as primary damage.

Dehydroascorbate (DHA)-reducing proteins were observed using a native-PAGE/activity staining method, and identified in soybean and rice by Western analysis using antibodies against homologous protein known to catalyse DHA reduction. Administration of 1 mM DHA apparently did not trigger DHA reductase activity in lupin and onion, whereas activity was increased in rice and barley.

This paper describes changes in leaf water status and in stem, petiole and leaf blade hydraulics preceding leaf senescence and shedding in Castanea sativa L. (chestnut). Measurements of maximum diurnal leaf conductance to water vapour (g_L), minimum water potential (ψ_L), hydraulic conductance per unit leaf surface area of stems (K_SL), petioles (K_PL) and leaf blades (K_LL) and number of functional conduits and inside diameter distribution were performed in June, September and October 1999. In September, still green leaves had undergone some dehydration as indicated by decreased g_L (by 75 %) and ψ_L with respect to June. In the same time, K_SL decreased by 88 %, while K_PL and K_LL decreased by 50 % and 20 % of the conduits of stems and 10 % of the petioles (all belonging to the widest diameter range) were no longer functioning, causing a decrease in the theoretical flow by 82 % in stems and 27 % in petioles. Stem xylem blockage was apparently due to tyloses growing into conduits. We advance the hypothesis that the entire process of leaf shedding and winter rest may be initiated by extensive stem embolism occurring during the summer.

Plant regeneration was achieved from the seed derived decotyledonated split embryo axes of six Indian cultivars of cotton (Gossypium hirsutum L.). Explants were cultured on Murashige and Skoog's basal medium supplemented with 2 % sucrose and 0.65 % agar. Incorporation of 0.25 % charcoal in the medium and incubation of the cultures at 30 ± 2 °C had synergistic effect on the frequency of shoot and root formation. The method employed is genotype independent, simple and rapid.

The eukaryotic unicellular microalgae Chlorella salina, Dicrateria inornata, and Isochrysis galbana were grown under control (fluorescent 20 W m^-2) and UV-B enhanced (UV-B^E, 0.5 W m^-2) fluorescent radiation. The growth rate showed marginal increase under UV-B^E. Decrease in protein content was observed in Dicrateria cells but in Chlorella an initial increase up to 4 d and in Isochrysis an increase at days 4 and 5 was noted. The chlorophyll a content showed marked increase in Chlorella and Isochrysis but in Dicrateria a decline was found. UV-B^E reduced the photosynthetic activity in all three species, but the reduction was larger in Chlorella and Dicrateria. Fluorescence excitation spectra for F₆₈₂ in Chlorella cells grown for 5 d under UV-B^E showed reduction in all peaks. In contrast to this, in Dicrateria and Isochrysis cells, the 530 and 590 nm excitation peaks increased with an appreciable decrease in the 466 nm peak. SDS-PAGE analysis revealed significant decrease in the contents of 47, 33, and 23 kDa polypeptides in Chlorella cells. In Dicrateria cells, significant loss in the content of 55, 38, and 18 kDa polypeptides was observed. The content of low molecular mass polypeptides (15 kDa) remained unaffected. Isochrysis cells were more stable in preserving the content of thylakoid polypeptides.

The effect of auxins [indole-3-butyric acid (IBA) and α-naphthaleneacetic acid (NAA)], phenolics (phloroglucinol and coumarin), a combination of auxins and phenolics, and a systemic fungicide (Bavistin, containing 50% carbendazim) on adventitious root formation in stem cuttings (current season's growth) of Cedrus deodara L. during winter and monsoon (rainy) seasons has been examined. Significant stimulation of rooting due to treatments was observed in cuttings planted in winter in the following order: 0.25 mM IBA (87.5% rooting) > 5 mM coumarin (70.8%) > IBA (0.5 mM) + coumarin (5 mM) (50.0%). In cuttings planted in monsoon only 0.05% Bavistin was found to be effective in inducing rooting (83.3%). Other treatments were ineffective and in some treatments drying of cuttings was noticed.

Diamine oxidase (DAO, EC 1.4.3.6.), which participates in oxidative catabolism of polyamines (PAs), was not detected in the dry viable chickpea (Cicer arietinum L.) seeds. From the time when the embryonic axis acquired an aerobic metabolism, DAO increased concomitantly with the growth of the embryonic axis and at the same time with the deterioration of the cotyledons, although in these organs the values were clearly lower than in the axis. The highest DAO activity in the embryonic axis of seedlings grown for 72 and 96 h was found in the elongation, differentiation and hypocotyl zones, while the lowest was in the apex and plumule. The absence of cotyledons promoted the early appearance of DAO in the embryonic axis. When germination occurred at supraoptimal temperatures (30 - 35 °C), DAO activity was sharply inhibited both in the cotyledons and in the embryonic axis. This inhibition was accentuated further in the presence of cyclohexylamine, an inhibitor of spermidine synthase activity, to such a degree that DAO was undetectable in the cotyledons. DAO inhibition by EGTA and the pronounced reversal induced by Ca²⁺ implies that calcium may be related to DAO activity. The presence of putrescine, spermidine and spermine in the germination medium stimulated DAO activity, although this activity was inhibited when the exogenous PA was cadaverine.

Greenhouse experiments were conducted in two years (1993-1994) with eggplants supplied with 1, 2 or 4 mM NH₄NO₃ as the N source in order to determine its influence on molybdenum (Mo) and nitrate (NO₃^-) content in leaf blades, petioles, and fruits as well as leaf nitrate reductase (NR) activity. The results reveal that 2 and 4 mM NH₄NO₃ altered shoot Mo distribution and thus affecting the NR activity.

To test whether different nitrogen form (nitrate or ammonium) in substrate can alter the response to elevated partial pressure of CO₂ (pCO₂) plants of perennial ryegrass (Lolium perenne cv. Bastion) were grown from seeds in growth chambers under pCO₂ of either 35 Pa (ambient, CA) or 70 Pa (elevated, CE) in a hydroponic system (with nutrient and pH control) for 24 d. Nitrogen was supplied as ammonium, nitrate or an equimolar mixture of both N forms. Under CE plants grew faster than their counterparts under CA during the first 14 d but after 23 d of cultivation stimulation disappeared. Despite the strong positive effect of mixed forms of N on plant growth, the beneficial effect of CE was similar to that in the other two N treatments. However, the almost alike final growth response to CE had different underlying mechanisms in different N treatments. Plants supplied with nitrate as a sole source of nitrogen had lower leaf mass ratio but much higher specific leaf area compared to plants supplied with ammonium. The decrease in the content of leaf organic N (per unit of structural dry mass) under CE was found only in leaves of plants supplied with ammonium on day 14. Nevertheless, the available form of N evidently contributes to changes of leaf N content under CE. The high levels of N and non-structural saccharides in plants supplied with ammonium at CE suggest that the CO₂ response of these plants was controlled by factors other than amount of available carbon and nitrogen.

Eryngium foetidum L. plants were regenerated from mature leaf and petiole explants through direct organogenesis without intervening callus phase. From leaf explants, adventitious multiple shoots raised on Murashige and Skoog (MS) medium supplemented with 4.43 μM benzylaminopurine (BAP) and 0.57 μM indole-3-acetic acid (IAA), whereas in petiole explants shoot regeneration occurred at 8.86 μM BAP and 0.57 μM IAAA. 80% of the leaf explants and 44% of petiole explants produced shoots after four weeks of culture. The regenerated plants were rooted on MS medium supplemented with 2.46 μM indole-3-butyric acid and 2.88 μM gibberellic acid. The plants were successfully established in the soil and showed 70.9% survival in the field.

The effects of 2-aminopurine, olomoucine, 6-dimethylaminopurine (inhibitors of cyclin-dependent kinases; CDK), and sodium vanadate (a potent inhibitor of protein phosphatases) on DNA endoreduplication were investigated during elongation and differentiation of the primary roots in Pisum sativum. When compared with the untreated control plants, at least one additional round of DNA replication was evidenced to occur within most cells, the majority of which have attained 4C DNA level, and a considerably greater portion of cells represented the endopolyploid state with nuclear DNA content approximating the 8C value. It is concluded that cellular commitment to DNA endoreduplication may appear not only as a consequence of suppression imposed directly upon CDK activity, but also as an indirect output connected with the decreased activity of cdc25 protein phosphatase, an enzyme necessary to turn the switch on for appropriate conformation of the CDK/cyclin B complex. By calculating the absorption profiles of Feulgen-stained nuclei, specific phosphorylation-dependent changes in chromatin condensation of endopolyploid cells have been revealed. It is proposed that acquisition of a certain critical level of chromatin condensation constitutes a prerequisite for additional rounds of DNA synthesis in plants.